神经母细胞瘤中Hsp27的表达及其与化疗药物关系的研究

Heat shock protein 27 expression in neuroblastoma and its relationship with chemotherapeutic agents

目的:探讨分析热休克蛋白27(heat shock protein 27,Hsp27)与儿童神经母细胞瘤临床病理参数及化疗药物的关系。方法:应用免疫组织化学(immunohistochemistry,IHC)法检测90例神经母细胞瘤(neuroblastoma,NB)、24例节细胞神经母细胞瘤(gan?glioneuroblastoma,GNB)和20例节细胞神经瘤(ganglioneuroma,GN)组织中Hsp27蛋白的表达;实时荧光定量PCR(fluorescence quantitative PCR,RT-qPCR)检测59例NB/GNB、5例GN组织标本,及人神经母细胞瘤SH-SY5Y细胞和耐顺铂SH-SY5Y细胞中Hsp27 mRNA的表达;应用IHC或PCR扩增测序法检测其中18例NB/GNB肿瘤标本化疗药物相关性分子靶标。结果:Hsp27蛋白在NB/GNB中阳性表达率为64.9%,明显高于GN的0(χ^2=28.994,P<0.05);在NB/GNB中,Hsp27阳性表达与肿瘤的分期、危险度、转移和预后有关(P<0.05),与患儿年龄、性别、Shimada组织学分型、N-myc有无扩增无关(P>0.05);Hsp27 mRNA在NB/GNB中的相对表达量为3.763±1.854,高于GN的2.476±0.698(P>0.05);Hsp27 mRNA在耐顺铂细胞中的表达明显高于亲代细胞(F=16.246,P<0.01)。检测化疗药物相关性分子靶标7个,结果显示Hsp27的表达与铂类药物敏感性相关。结论:本研究初步显示Hsp27的表达与NB的分期、危险度、转移、预后相关,可能参与了NB/GNB的发生发展转移。Hsp27的表达与铂类敏感性相关,需进一步增大样本量来验证其相关性。

Objective:To investigate the expression of heat shock protein 27(Hsp27)in children with neuroblastomas and analyze the relationship of Hsp27 expression and clinicopathological parameters with chemotherapeutic agents.Methods:Immunohistochemical(IHC)staining was performed to detect the expression of Hsp27 protein in 90 neuroblastomas(NBs),24 ganglioneuroblastomas(GNBs),and 20 ganglioneuromas(GNs).Quantitative real-time polymerase chain reaction PCR(RT-qPCR)examinations were performed to detect the expression of mRNA in 59 NBs/GNBs,5GNs,neuroblastoma cell lines,and drug-resistant cell lines.Immunohistochemical staining,PCR amplification,and sequencing were used to examine the expression or mutation of drug-related genomic markers in 18 NBs/GNBs.Results:The positive expression rates of Hsp27 in NBs/GNBs were significantly higher than that in GNs(64.9%vs.0,χ^2=28.994,P<0.05).In NBs/GNBs,positive Hsp27 expression was associated with tumor stage,risk stratification,metastasis,and prognosis(P<0.05),but was not associated with age,sex,Shimada classification,or N-MYC(P>0.05).The mRNA levels of Hsp27 in NBs/GNBs were higher than those in GNs(3.763±1.854 vs.2.476±0.698,P>0.05).The mRNA levels of Hsp27 in the drug-resistant cell line were significantly higher than those in the neuroblastoma cell line(F=16.246,P<0.01).Seven genomic markers of anti-neoplastic agents were examined,and Hsp27 expression was found to be related to platinum resistance.Conclusions:Hsp27 expression was associated with tumor stage,risk stratification,metastasis,and prognosis.Therefore,Hsp27 may promote tumorigenesis,progression,and metastasis.Furthermore,Hsp27 expression is related to platinum resistance.Further studies with larger sample sizes areneeded to verify these correlations.